目的:本研究旨在建立双抗原夹心ELISA(DAgS-ELISA)方法,准确,并定量检测抗白念珠菌烯醇酶1(CaEno1)的总抗体,以诊断侵袭性念珠菌病(IC)。
方法:使用重组CaEnol和单克隆抗体作为标准品开发DAgS-ELISA。性能评估包括检测限,准确度,和可重复性。使用DAgS-ELISA分析来自全身性念珠菌病小鼠的血清中抗CaEnol抗体水平的动态变化。来自IC的患者血清样本,念珠菌定植,细菌感染,和健康对照组用DAgS-ELISA和间接ELISA进行分析。
结果:DAgS-ELISA在线性范围和测试背景方面优于间接ELISA。在全身性念珠菌病小鼠中,在动态抗体水平中观察到独特的“双峰”模式。此外,通过DAgS-ELISA和间接ELISA检测到的CaEno1抗体阳性率具有较高的一致性.虽然患者组的阳性率不同,在各阳性患者组中未检测到抗体水平的显著差异.
结论:DAgS-ELISA为IC诊断提供了一种可靠的新方法,快速启用,准确,并定量检测CaEno1抗体。其临床应用和有效性需要进一步验证和优化。
OBJECTIVE: This study aimed to develop a double antigen sandwich ELISA (DAgS-ELISA) method for more efficient, accurate, and quantitative detection of total antibodies against Candida albicans enolase1 (CaEno1) for diagnosing invasive candidiasis (IC).
METHODS: DAgS-ELISA was developed using recombinant CaEno1 and a monoclonal antibody as the standard. Performance evaluation included limit of detection, accuracy, and repeatability. Dynamic changes in antibody levels against CaEno1 in serum from systemic candidiasis mice were analyzed using DAgS-ELISA. Patient serum samples from IC, Candida colonization, bacterial infections, and healthy controls were analyzed with DAgS-ELISA and indirect ELISA.
RESULTS: DAgS-ELISA outperformed indirect ELISA in terms of linear range and test background. In systemic candidiasis mice, a distinctive \'double-peak\' pattern in dynamic antibody levels was observed. Additionally, there was a high level of consistency in the positive rates of CaEno1 antibodies detected by both DAgS-ELISA and indirect ELISA. While the positivity rates differed among patient groups, no significant variations in antibody levels were detected among the various positive patient groups.
CONCLUSIONS: DAgS-ELISA offers a reliable novel approach for IC diagnosis, enabling rapid, accurate, and quantitative detection of CaEno1 antibodies. Further validation and optimization are needed for its clinical application and effectiveness.